Abstract
DNA genotyping has traditionally been performed by slab-gel electrophoresis. The method is well established, very effective and reliable, but inherently slow and labor intensive. Capillary gel electrophoresis is now becoming recognized as an important alternative to slabs since it offers higher throughput and automation (1). Nevertheless, the fast growing need for DNA analysis capacity due, for example, to the sequencing of entire genomes and the establishment of complex DNA data banks, seems to demand even more powerful DNA analysis tools. Microdevice electrophoresis is being increasingly explored since it may allow electrophoretic DNA analysis approaching the theoretical performance limits of the method due to unique sample loading characteristics and the employment of very short separation distances (2). It has already been demonstrated that genotyping can be performed 10-100 faster than on capillaries and slabs (see Subheading 3.4.). In addition, the fabrication and operation of high-density electrophoretic microdevices for high-sample throughput should be straightforward. Microfabrication might also permit the total integration of entire sample processing sequences (e.g., PCR, sample cleanup, separation) on one single device, potentially leading to drastically decreased sample and reagent volumes, significantly less human interference, and increased speed of analysis (3).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Mullikin, J. C., and McMurray, A. A. (1999) Sequencing the genome fast. Science 283, 1867–1868.
Freemantle, M. (1999) Downsizing chemistry. Chemical analysis and synthesis on microchips promise a variety of potential benefits. Chem. Eng. News 22, 27–36.
Service, R. F. (1998) Coming soon: the pocket DNA sequencer. Science 282, 399–401.
Harrison, D. J., Fluri, K., Seiler, K., Fan, Z. Effenhauser, C. S., and Manz, A. (1993) Micromachining a miniaturized capillary electrophoresis-based chemical analysis system on a chip. Science 261, 895–897.
Simpson, P. C., Wooley, A. T., and Mathies, R. A. (1998) Microfabrication technology for the production of capillary array electrophoresis chips. Biomed. Microdevices 1, 7–25.
Simpson, P. C., Roach, D., Woolley, A. T., Thorsen, T., Johnston, R., Sensabaugh, G. F., et al. (1998) High-throughput genetic analysis using microfabricated 96-sample capillary array electrophoresis microplates. Proc. Natl. Acad. Sci. USA 95, 2256–2261.
Schmalzing, D., Koutny, L., Adourian, A., Belgrader, P. H., Matsudaira, P., and Ehrlich, D. (1997) DNA typing in thirty seconds with a microfabricated device. Proc. Natl. Acad. Sci. USA 94, 10,273–10,278.
Schmalzing, D., Koutny, L., Chisholm, D., Adourian, A., Matsudaira, P., and Ehrlich, D. (1999) Two-color multiplexed analysis of eight short tandem repeat loci with an electrophoretic microdevice. Anal. Biochem. 270, 148–152.
Schmalzing, D., Koutny, L., Adourian, A., Matsudaira, P., and Ehrlich, D. (1997) Ultrafast STR analysis by microchip gel electrophoresis. In: Proceedings from the 8th International Symposium on Human Identification. Promega Corp., pp. 112–118.
Jacobson, S. C., and Ramsey, J. M. (1996) Integrated microdevice for DNA restriction fragment analysis. Anal. Chem. 68, 720–723.
Xie, W., Yang, R., Xu, J., Zhang, L., Xing, W., and Cheng, J. (2001) Microchip-based capillary electrophoresis systems, in Capillary Electrophoresis of Nucleic Acids, Vol. 1 (Mitchelson, K. R., and Cheng, J., eds.), Humana Press, Totowa, NJ, pp. 67–83.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc.
About this protocol
Cite this protocol
Schmalzing, D., Koutny, L., Adourian, A., Chisholm, D., Matsudaira, P., Ehrlich, D. (2001). Genotyping by Microdevice Electrophoresis. In: Mitchelson, K.R., Cheng, J. (eds) Capillary Electrophoresis of Nucleic Acids. Methods in Molecular Biology™, vol 163. Humana Press. https://doi.org/10.1385/1-59259-116-7:163
Download citation
DOI: https://doi.org/10.1385/1-59259-116-7:163
Publisher Name: Humana Press
Print ISBN: 978-0-89603-765-6
Online ISBN: 978-1-59259-116-9
eBook Packages: Springer Protocols